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Objective:To explore the prediction model of tissue chip technology for the chemotherapy response of patients with colorectal cancer.Methods:217 patients with colorectal cancer who had received standardized chemotherapy in the Affiliated People’s Hospital of Ningbo University from Jan. 2017 to Dec. 2019 were prospectively selected. The patients were randomly divided into training set (152 cases) and test set (65 cases) according to the ratio of 7:3, and were followed up for 6 months. The clinical data of the patients in the training set were compared, the expression levels of Ang-2, caspase-3 and CD147 in the patients were analyzed by tissue microarray technology, and the related factors affecting the responsiveness of colorectal cancer chemotherapy were analyzed by the Logistic regression model. R software was used based on the training set. A nomogram prediction model was built and model performance on the test set was evaluated.Results:One case was excluded from the training center, and 151 cases were finally included, including 93 cases in the chemotherapy response group and 58 cases in the chemotherapy response group. The tumor diameter, serum carcinoembryonic antigen, caspase3, Ang2 expression level, and the proportion of clinical stage IV in the poor chemotherapy group were significantly higher than those in the good chemotherapy group (all P<0.05) ; Logistic regression showed tumor diameter ( OR=2.394), serum carcinoembryonic antigen ( OR=1.878), caspase-3 ( OR=4.261), Ang-2 expression level ( OR=5.457), and clinical stage IV ( OR=5.954) were independent risk factors for adverse drug reactions in patients with colorectal cancer (all P<0.05). The consistency index (C-index) for predicting the factors related to adverse chemotherapy reactions in patients with colorectal cancer was 0.915. External verification showed that the sensitivity was 86.96%, the specificity was 92.50%, and the accuracy was 90.48% (42/65) . Conclusion:The expression levels of Ang-2 and caspase-3 are correlated with the responsiveness of colorectal cancer to chemotherapy, and can be used as predictive indicators to evaluate the responsiveness of colorectal cancer to chemotherapy.
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@#[Abstract] Objective: To investigate the expression of chemokine-like factor-like MARVEL transmembrane domain-containing family member 6 (CMTM6) in breast cancer tissues and its correlation with clinicopathological features and prognosis of patients. Methods:Atotal of 136 breast cancer tissue chips (purchased from Superchip Company), including 42 pairs of matched cancer and paracancerous tissues, were used for this study. The expression level of CMTM6 in cancer and paracancerous tissues was detected by immunohistochemistry. The comparison of CMTM6 expression between breast cancer and paracancerous tissues was conducted by paired χ2 test. The relationship between CMTM6 expression in breast cancer tissues and the clinicopathological characteristics of patients was analyzed by χ2 test. Kaplan-Meier and Log rank test analyses were used to analyze the relationship between CMTM6 expression and the survival of patients, and Cox model was used to evaluate the effect of different indicators on the prognosis of patients. Results: The expression of CMTM6 in breast cancer tissues was significantly higher than that in paracancerous tissues (P<0.01). The expression of CMTM6 was correlated with pathological type of breast cancer and HER2 positivity (P<0.05). The survival time of patients in CMTM6 high expression group was significantly shorter than that of patients in CMTM6 low expression group (P<0.05). Pathological type (HR=10.374, 95%CI: 3.529-30.497, P<0.01), TNM stage (HR=4.599, 95%CI: 1.784-11.856, P<0.01), triple-negative breast cancer (HR=3.370, 95%CI: 1.055-10.761, P<0.05) and high expression of CMTM6 (HR=0.195, 95%CI: 0.073-0.518, P<0.01) were independent risk factors for prognosis of breast cancer patients. Conclusion: CMTM6 is highly expressed in breast cancer tissues, which can be used as a risk factor for prognosis evaluation of breast cancer patients.
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Purpose To study the expression of TMPRSS2, ERG and ETV1 in prostatic cancer and their clinical pathologic signifi-cance. Methods Tissue microarray and immunohistochemistry (MaxVision) were used to detect TMPRSS2, ERG and ETV1 expres-sion in 70 prostatic cancer tissues, 10 prostatic intraepithelial neoplasia tissues and 18 benign prostate tissues. Results There was no statistical significance on positive rate of the expression of TMPRSS2 among prostatic cancer tissues, prostatic intraepithelial neoplasia tissues and benign prostate tissues (P>0. 05). The positive rate (81. 4%) of ERG in prostatic cancer tissues was significantly higher than that in prostatic intraepithelial neoplasia tissues ( 30. 0%) and benign prostate tissues ( 0. 0 ) ( P 0. 05). The expression of TMPRSS2, ERG and ETV1 was positively correlated to Gleason score and clinical stage (P0. 05). Conclusion ERG and ETV1 are expected to become therapeutic targets for prostate cancer. Detecting TMPRSS2, ERG and ETV1 at the same time is helpful to diagnosis and differential diagnosis of prostatic cancer, which might be new molecule markers of prostate cancer.
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Objective: To explore the clinical significance of transcription factor Sp1 expression in hepatocellular carcinoma (HCC) and association between Sp1 expression and survival in HCC patients. Methods:With the use of immunoreactivity, Sp1 expres-sion and its correlation with other clinicopathological characteristics were examined in a tissue microarray that contains samples from 98 HCC patients. Results:HCC tissues expressed markedly higher levels of Sp1 than did adjacent normal liver tissues. Sp1 expression was closely associated with microvascular invasion, which suggests that HCC with more microvascular invasion is prone to have in-creased Sp1 expression. Overexpression of Sp1 correlates with significantly shorter overall survival and higher recurrence rates in HCC patients after curative resection. Conclusion:Transcription factor Sp1 may be an independent prognostic factor for both overall surviv-al and cumulative recurrence rate.
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Objective To study the expression of forkhead box 2(FOXC2)and delta-like liGand 4 (DLL4)in invasive ductal carcinoma(IDC)of breast and the clinical siGnificance. Methods The expression of FOXC2 and DLL4 in l22 cases of IDC(Grade Ⅰ33,Grade Ⅱ40,GradeⅢ 49)were observed by tissue chip and immunohistochemistry. The relationship of the expression with clinicopatholoGical characteristics and between FOXC2 and DLL4 were statistically analyzed. PCR experiment was performed in normal breast tissue,ductal carcinoma in situ(DCIS)IDC Grade Ⅰ,Grade Ⅱ and Grade Ⅲ(l0 cases respectively). Results The positive rate of FOXC2 and DLL4 in IDC was 77. 87% and 74. 59% respectively. A hiGher expression was observed in GradeⅢ than in GradeⅠand Ⅱ( P<0. 05 ). The expression of FOXC2 was related to the neGative expression of ER. The expression of DLL4 was related to the tumour size,clinical staGe and lymph node metastasis( P<0. 05). The RCR of FOXC2 and DLL4 were Gradually increased in normal breast tissue,DCIS,IDC GradeⅠ, Grade Ⅱ and Grade Ⅲ(P<0. 05). Moreover the expression of FOXC2 was related to the expression of DLL4(r=0. 233,P=0. 0l0). Conclusion FOXC2 and DLL4 miGht toGether have influence on the proGression and outcome of breast carcinoma,and could be important markers of proGnosis. DLL4 miGht be reGulated by variety of factors includinG FOXC2 at the same time.
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The tissue chip is a special tissue section,which has several tens to thousands mini tissues of regular arrangement on the one glass slide.According to the deficiencies in experimental teaching section of morphology at present,the author introduced the characteristics and values of the tissue chip and explored its application prospect in experimental teaching of morphology.
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Objective To detect the expression of KiSS-1, KiSS-1R and MMP-9 in hepatocellular carcinoma (HCC). To study the correlation of KiSS-1, KiSS-1R and MMP-9 expression with invasion and metastasis of HCC, and to explore the underlying mechanisms. Methods The expression of KiSS-1 , KiSS-1R mRNA in 33 HCC samples, 26 non-neoplastic adjacent liver tissue samples and 13 non-neoplastic distant liver tissue samples were detected by RT-PCR. Tissue chips were constructed by modified manual tools, which contained HCC, non-neoplastic adjacent liver tissues, non-neoplastic distant liver tissues, normal liver tissues and intrahepatic metastasis lesions. The expression of KiSS-1 and MMP-9 protein was determined by tissue chips, immunohistochemistry and semi-quantitative image analysis in 150 HCC, 137 non-neoplastic adjacent liver tissue, 98 non-neoplastic distant liver tissues, 16 normal liver tissues and 37 intrahepatic metastasis lesion samples. Results The results of RT-PCR showed that compared with the non-neoplastic adjacent liver tissues and the non-neoplastic distant liver tissues, the expression of KiSS-1 mRNA in HCC was significantly lower (P＜0.01). The expression of KiSS-1R mRNA did not changed in HCC and non-neoplastic liver tissues (P＞0.05). The expression of KiSS-1 protein was lower in HCC with metastasis and in clinical stage Ⅲ than that in those with non-metastasis, and in clinical stages Ⅰ and Ⅱ . It was also higher in the primary than in the metastasis lesions (P＜0.01, respectively). The expression of MMP-9 was higher in tumors having peplos invasion and metastasis than in those with negative peplos invasion and non-metastasis. It was lower in the primary than the metastasis lesions (P＜0. 01, respectively).Negative correlation between KiSS-1 and MMP-9 expression was found in HCC(r=- 0.340,P＜0.01). Conclusions The imbalance between KiSS-1 and MMP-9 expression might play an important role in enhancing the invasive and metastatic capacity of HCC. Loss of KiSS-1 expression might predict an aggressive clinical behavior and was associated with metastatic potential in HCC.
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Objective: To prepare rabbit polyclonal antibody against human P-element-induced wimpy testis like 2 (HIWI2) protein, identify its properties and investigate its distribution in normal and tumor tissues by means of tissue chip. Methods: PIWIL2 peptide was synthesized chemically and conjugated to Keyhole limpet hemocyanin (KLH) as an immunogen. Then the PIWIL2-KLH conjugation was injected into rabbits subcutaneously to produce polyclonal antibodies. The specificity and sensitivity of antibodies were identified by ELISA and Western blotting after purification by affinity chromatography. PIWIL2 was then immuno-stained on the tissue chip to study its distribution in the normal and tumor tissues. Results: Rabbit's antibodies against human PIWIL2 were prepared after the injection of PIWIL2-KLH conjugation subcutaneously. These antibodies were identified as PIWIL2 peptides by ELISA and Western blotting assay. PIWIL2 protein expression was tissue-specific in tumor tissues, with PIWIL2 protein found in the cytoplasm of smooth muscle cells of most normal and tumor tissues. Conclusion: The polyclonal antibodies against human PIWIL2 protein have been successfully prepared, which provides a basis for further study on the role of PIWIL2 in the pathway of miRNA/RNA.
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potential use in clinical therapy and prognosis estimation of the tumor.
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Objective To study the expression of tyrosine protein kinase in hepatocellular carcinoma(HCC)and observe the correlation between tyrosine protein kinase and clinical features.Methods A total of 30 cases with HCC were enrolled in this study.ERK,P38,C-jun,JAK2,STAT3 and STAT5 were detected by immunohistochemical method using tissue chip technology.Results The expression of ERK(0.220±0.033),P38(0.174±0.024),C-jun(0.183±0.064),JAK2(0.192±0.044),STAT3(0.197±0.078)and sTAT5(0.181±0.066)in HCC was significantly higher than that(0.065±0.028,0.058±0.028,0.042±0.016,0.070±0.030,0.052±0.024,0.052±0.023)in cirrhosis 1iver tissues(P＜0.01).There was significantly positive correlation of the expression between ERK,C-jun,JAK2,STAT3 and STAT5(P＜0.01 or P＜0.05).But the expression of P38 was negatively correlated with ERK in the HCC tissues(r=-0.404,P＜0.05).JAK2 had significant correlation with tumor differentiation.The expression of J AKz in stage Ⅲ was significantly higher than that in stage Ⅰ and Ⅱ cancer tissues.Conclusion There is important significance of the excessive activation of MAPK and JAK-STAT signaI transduction in hepatocellular carcinoma process.The unbalance of signal transduction might be one of the pathogenesis of tumor progress.
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Objective To study the difference of IGF-1,Bcl-2,survivin and Ki-67 protein expression in reactive and neoplastic astrocytes and the significance of them.Methods Immunohistochemistry and tissue microarray techniques were used to determinate the expression of IGF-1,Bcl-2,survivin and Ki-67 protein in normal brain tissues, astrocytes proliferation,low-grade astrocytomas, and high-grade astrocytomas.Results The expression of IGF-1,Bcl-2,survivin and Ki-67 protein were all negative in control group.The positive expression rates of them in reactive astrocytes were 28.9%, 26.7%, 26.7% and 22.2%,respectvely; and in low-grade astrocytomas were 63.8%, 50.0%, 53.2%,70.2%; in high-grade astrocytomas were 88.9%, 79.2%, 88.1%,95.2%.IGF-1,Bcl-2,survivin and Ki-67 all had significant difference among the three experimental groups(P
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Objective To investigate the expression and clinical significance of Bmi-1,Podoplanin and p53 in human thymic epithelial tumor(TET) according to new histological classification of World Health Organization. Methods One hundred and ten TET tissue chips were prepared,and the expressions of Bmi-1,Podoplanin and p53 were detected by NovoLinkTM Polymer detection systems. Results The positive expressions of Bmi-1,Podoplanin and p53 were 68.2%,41.8% and 42.7%,respectively.The positive expressions of Bmi-1,Podoplanin and p53 were significantly associated with pathologic histological classification,Masaoka staging and clinic pathological grading(P
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Objective To prepare polyclonal antibodies against human PIWIL and to identify their property and tissue distribution of PIWIL.Methods PIWIL polypeptide was synthesized and conjugated to Keyhole limpet hemocyanin(KLH) as an immunogen.Then PIWIL-KLH conjugations were injected into rabbits subcutaneously to produce polyclonal antibodies.The specificity and sensitivity of antibodies were identified by ELISA and Western blot after purification by affinity chromatography.PIWIL were then stained on the tissue chip to study their distribution.Results Rabbit antibodies against PIWIL were prepared after injection of PIWIL-KLH conjugation.These antibodies specially recognized PIWIL peptides.Expression of PIWIL was found in the cytoplasm of epithelia cells of varied normal tissues and tumor tissues.Conclusion The successful preparation of the polyclonal antibody against PIWIL will provide an efficient reagent for further study of its role in the pathway of miRNA and RNA interference and in the pathogenesis of human disease.
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Objective To detect the expression and potential correlation of ERK1 and P16 in gastrointestinal stromal tumor group and in control group.Methods Tissue chip and immunohistochemistry Elivison were used to detect the expression of ERK1 and P16 in 40 cases of gastrointestinal stromal tumor and 40 cases of control tissues.Quantitative analysis of mean absorbent density of the expression of ERK1 and P16 was conducted with image analytic software.Results The expressions of ERK was higher in GIST group than that in control group(P
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【Objective】To establish a simple method for preparing tissue chip.【Methods】One hundred and thirty mammary cancer and intestinal cancer specimens were fixed by formaldehyde(40?g/L) and embedded by wax,and then were made into routine tissue slices and stained by haematoxylin and eosin(HE).Wax slices with tissue chip were prepared by fixing a tissue slice(0.1 cm in diameter)into the hole(0.2 cm in diameter and 2～4mm in depth) of a blank wax slice and were sliced into 4?m-thick slices.The tissue chips were mounted on the slide and stained by immunohistochemistry after treatment.【Results】A slide held 45 tissue chips and 130 specimens could be all detected only on 3 slides,which saved time and the amount of reagent.The immunohistochemical results showed that the positive location was accurate and the background was clear.【Conclusion】This method for preparing tissue chip is simple and effective,and can save the amount of reagent.It is easy for the comparative and separated detection of specimens.
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Objective To investigate the correlation of ? catenin expression with the carcinogenesis and progression of human colorectal cancer Methods The colorectal adenoma carcinoma tissue chips including 400 cores were constructed and the expression of ? catenin in 220 cases of colorectal adenoma and carcinoma was detected by immunohistochemical staining Results The effective rates of information by tissue chips stained by hematoxylin eosin and immunohistochemistry were 98 8% and 98 0%, respectively The nuclear expression rate of ? catenin was 35 9% in malignant changes in adenoma, which was significantly higher than that in adenoma (16 7%) and carcinoma (19 7%) ( P
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Purpose:Constructing a high-flux tissue microarray/tissue chip and detecting IGF-Ⅱprotein expression of cases by it, and to determine the correlation between IGF-Ⅱprotein expression and lung cancer. Methods:A series of tissue chips were prepared by using tissue arrayer with samples from lung cancers of different histological classifications. Specimens from 54 cases of lung cancer and 10 cases of normal lung tissues were detected immunohistochemically on a tissue chip for IGF-Ⅱprotein expression and its correlation to clinic-pathological parameters was analyzed statistically. Results:Positive rate of IGF-Ⅱprotein in lung cancer was 42.6%(23/54), which was higher than that of normal lung(0.0%, 0/10, P0.05). Conclusions:IGF-Ⅱprotein might be related to the malignant behaviors of lung cancer. Detecting the expression of IGF-Ⅱ protein probably can predict the prognosis of lung cancer. It is feasible to utilize tissue chip for screening of clinical tissue specimens on a large scale.
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0.05).?-defensin 2 was detected in the specimens of vocal cord polyp,but very little in the subjects of other two groups.Its expression level was significantly higher in the vocal cord polyp than that of the other two groups(P
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Objective To study the significance of expression of hypoxia-inducible factor 1 alpha(HIF-1?) in gastric mucosal diseases. Methods Using tissue chip technique to creat tissue chip in 120 cases of vary gastric mucosal disease,and using S-P immunohistochemical methods to detect the expressions of HIF-1? in these tissue chips. Results The positive rates of HIF-1? was 28.3% in 120 tissue chips of gastric mucosal diseases. The positive rates of HIF-1? was 0, 10.0%, 40.0% and 63.3% in chronic superficial gastritis, chronic atrophic gastritis with intestinal metaplasia, chronic atrophic gastritis with dysplasia and intestinal type gastric cancer, respectively. The positive rates of HIF-1? of intestinal type gastric cancer and chronic atrophic gastritis with dysplasia were significantly higher than that in chronic superficial gastritis and chronic atrophic gastritis with intestinal metaplasia(P
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Objective:To study the expressions of Skp2 and P27 in the normal and tumor tissues of salivary glands,and then to evaluate their significance in the development and progression of salivary tumor.Methods:SP immunohistochemical method was used to evaluate the expression of Skp2 and P27 in the normal and tumor tissues of salivary glands. Results:The rates of Skp2 and P27 expression were neither significantly different between polymorphic adenoma and basal cell adenomas, nor significantly different between adenoid cystic carcinoma and mucoepidemoid carcinoma. The rates of Skp2 positive were significantly different between normal salivary glands and benign salivary tumor, as well as between normal salivary glands and malignant salivary tumor. The rates of Skp2 and P27 positive were significantly different between normal salivary glands and malignant salivary tumor.Conclusion:Skp2 overexpression and decreased expression of P27 might play an important role in the development and progression of salivary tumor, overexpression of Skp2 protein may lead to increase of P27 degradation.